PurposeExpresses 3xFLAG-dCas9 in E. coli in a tetracycline-dependent manner for enChIP analysis to purify specific genomic regions of interest.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||64325||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonep15A vector
- Backbone size w/o insert (bp) 2598
- Total vector size (bp) 6777
Vector typeBacterial Expression, CRISPR ; Tetracycline-inducible
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberLow Copy
SpeciesSynthetic; S. pyogenes
Insert Size (bp)4179
- Promoter pLtetO-1
/ Fusion Protein
- 3xFLAG (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site Bgl II (not destroyed)
- 3′ cloning site Xho I (not destroyed)
- 5′ sequencing primer cgattccgacctcattaagcagc
- 3′ sequencing primer tgcctggagatccttactcgaGTTAGTCACCTCCTAGCTGACTCAAATCAAT (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byA portion of this plasmid was derived from a plasmid made by Stanley Qi (Addgene plasmid 44249)
Terms and Licenses
This plasmid is compatible with gRNA expression plasmids for E. coli, such as Addgene Plasmid #44251 ( http://www.addgene.org/44251 ).
For more information on Fujii Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/fujii/
Plasmid also described in Fujita, T. and Fujii, H., Biochem Biophys Res Commun. 2013 Sep 13;439(1):132-6. doi: 10.1016/j.bbrc.2013.08.013, PMID: 23942116
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:3xFLAG-dCas9/p-bacteria was a gift from Hodaka Fujii (Addgene plasmid # 64325 ; http://n2t.net/addgene:64325 ; RRID:Addgene_64325)
For your References section:An enChIP system for the analysis of bacterial genome functions. Fujita T, Yuno M, Fujii H. BMC Res Notes. 2018 Jun 14;11(1):387. doi: 10.1186/s13104-018-3486-3. 10.1186/s13104-018-3486-3 PubMed 29898790